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Article

Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy

1
Adolphe Merkle Institute, University of Fribourg, Chemin des Verdiers 4, 1700 Fribourg, Switzerland
2
Department of Life Sciences, Nano for Environment Unit, Water Quality Group, Av. Mestre José Veiga s/n, 4715-330 Braga, Portugal
3
In Vitro Toxicology Group, Swansea University Medical School, Swansea SA2 8PP, Wales, UK
4
Health Effects and Exposure Science, Pacific Northwest National Laboratory, Richland, WA 99352, USA
5
PETA International Science Consortium Ltd., London N1 9RL, UK
6
Department of Chemistry, University of Fribourg, Chemin du Musée 9, 1700 Fribourg, Switzerland
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Nanomaterials 2019, 9(12), 1765; https://doi.org/10.3390/nano9121765
Received: 14 November 2019 / Revised: 2 December 2019 / Accepted: 7 December 2019 / Published: 11 December 2019
(This article belongs to the Section Biology and Medicines)
The overt hazard of carbon nanotubes (CNTs) is often assessed using in vitro methods, but determining a dose–response relationship is still a challenge due to the analytical difficulty of quantifying the dose delivered to cells. An approach to accurately quantify CNT doses for submerged in vitro adherent cell culture systems using UV-VIS-near-infrared (NIR) spectroscopy is provided here. Two types of multi-walled CNTs (MWCNTs), Mitsui-7 and Nanocyl, which are dispersed in protein rich cell culture media, are studied as tested materials. Post 48 h of CNT incubation, the cellular fractions are subjected to microwave-assisted acid digestion/oxidation treatment, which eliminates biological matrix interference and improves CNT colloidal stability. The retrieved oxidized CNTs are analyzed and quantified using UV-VIS-NIR spectroscopy. In vitro imaging and quantification data in the presence of human lung epithelial cells (A549) confirm that up to 85% of Mitsui-7 and 48% for Nanocyl sediment interact (either through internalization or adherence) with cells during the 48 h of incubation. This finding is further confirmed using a sedimentation approach to estimate the delivered dose by measuring the depletion profile of the CNTs. View Full-Text
Keywords: adherent cell cultures; carbon nanotubes; depletion; quantification; sedimentation; UV-VIS-NIR spectroscopy adherent cell cultures; carbon nanotubes; depletion; quantification; sedimentation; UV-VIS-NIR spectroscopy
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MDPI and ACS Style

Septiadi, D.; Rodriguez-Lorenzo, L.; Balog, S.; Spuch-Calvar, M.; Spiaggia, G.; Taladriz-Blanco, P.; Barosova, H.; Chortarea, S.; Clift, M.J.D.; Teeguarden, J.; Sharma, M.; Petri-Fink, A.; Rothen-Rutishauser, B. Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy. Nanomaterials 2019, 9, 1765. https://doi.org/10.3390/nano9121765

AMA Style

Septiadi D, Rodriguez-Lorenzo L, Balog S, Spuch-Calvar M, Spiaggia G, Taladriz-Blanco P, Barosova H, Chortarea S, Clift MJD, Teeguarden J, Sharma M, Petri-Fink A, Rothen-Rutishauser B. Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy. Nanomaterials. 2019; 9(12):1765. https://doi.org/10.3390/nano9121765

Chicago/Turabian Style

Septiadi, Dedy, Laura Rodriguez-Lorenzo, Sandor Balog, Miguel Spuch-Calvar, Giovanni Spiaggia, Patricia Taladriz-Blanco, Hana Barosova, Savvina Chortarea, Martin J.D. Clift, Justin Teeguarden, Monita Sharma, Alke Petri-Fink, and Barbara Rothen-Rutishauser. 2019. "Quantification of Carbon Nanotube Doses in Adherent Cell Culture Assays Using UV-VIS-NIR Spectroscopy" Nanomaterials 9, no. 12: 1765. https://doi.org/10.3390/nano9121765

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